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Odontology

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Genetic polymorphisms influence gene expression of human periodontal ligament fibroblasts in the early phases of orthodontic tooth movement

  • Erika Calvano Küchler
  • Agnes Schröder
  • Paola Corso
  • Rafaela Scariot
  • Gerrit Spanier
  • Peter Proff
  • Christian KirschneckEmail author
Original Article
  • 68 Downloads

Abstract

Genetic polymorphisms could be involved in the individual rate of OTM (orthodontic tooth movement) corresponding to the clinical phenomenon of “slow movers” and “fast movers”. This study evaluated, if genetic polymorphisms in RANK, RANKL, OPG, COX2 and IL6 are associated with the expression of RANKL, OPG, COX2 and IL6 by human periodontal ligament (hPDL) fibroblasts during OTM. Primary hPDL fibroblasts from periodontal connective tissue of teeth extracted from 57 human subjects for medical reasons were collected, isolated, cultivated and characterized. To simulate orthodontic forces in PDL pressure areas, a physiological compressive force of 2 g/cm2 was applied to the hPDL fibroblasts under cell culture conditions at 70% confluency for 48 h, using a glass disc. Thereafter we analysed relative expression of RANKL, OPG, COX2 and IL6 by RT-qPCR. We also performed genotyping analysis of seven genetic polymorphisms in RANK, RANKL, OPG, COX2 and IL6. Relative gene expression was compared among the genotypes. The genotype TT in polymorphism rs9594738 (RANKL) had a higher RANKL expression in the recessive model (p = 0.021; TT vs. CT + CC). For polymorphism rs9594738 (RANKL), in the recessive model, TT was associated with a higher RANKL/OPG expression ratio (p = 0.013; TT vs. CT + CC). In the dominant model, GG genotype in rs5275 (COX2) was associated with a lower gene expression of COX2 (p = 0.04; GG vs. AA + AG). Genetic polymorphisms in genes associated with OTM affect the relative force-induced upregulation of these genes in hPDL fibroblasts.

Keywords

Genetic polymorphism Orthodontics Periodontal ligament Bone Connective tissue 

Notes

Funding

This study was funded by the Bavarian University Centre for Latin America BAYLAT (Grant ID: Kirschneck 12/2017) and by the São Paulo Research Foundation (FAPESP) (Grant ID: Küchler 2015/06866-5).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Approval for the collection and usage of hPDL fibroblasts was obtained from the ethics committee of the University of Regensburg, Germany [approval number 12-170-0150]. This article does not contain any studies with animals.

Supplementary material

10266_2019_475_MOESM1_ESM.pdf (324 kb)
Supplementary material 1 Raw genotype and phenotype gene expression data of all individual subjects investigated (PDF 323 kb)

References

  1. 1.
    Graber TM, editor. Orthodontics: current principles and techniques. 4th ed. St. Louis: Elsevier Mosby; 2005.Google Scholar
  2. 2.
    Meikle MC. The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt. Eur J Orthod. 2006;28:221–40.CrossRefGoogle Scholar
  3. 3.
    Kanzaki H, Chiba M, Shimizu Y, Mitani H. Periodontal ligament cells under mechanical stress induce osteoclastogenesis by receptor activator of nuclear factor kappaB ligand up-regulation via prostaglandin E2 synthesis. J Bone Miner Res. 2002;17:210–20.CrossRefGoogle Scholar
  4. 4.
    Schröder A, Bauer K, Spanier G, Proff P, Wolf M, Kirschneck C. Expression kinetics of human periodontal ligament fibroblasts in the early phases of orthodontic tooth movement. J Orofac Orthop. 2018;79:337–51.CrossRefGoogle Scholar
  5. 5.
    Ren Y, Maltha JC, Kuijpers-Jagtman AM. Optimum force magnitude for orthodontic tooth movement: a systematic literature review. Angle Orthod. 2003;73:86–92.PubMedGoogle Scholar
  6. 6.
    Proffit WR, editor. Biologic basis of orthodontic therapy. In: Contemporary orthodontics. 4th ed. St. Louis: Elsevier Mosby; 2007.Google Scholar
  7. 7.
    Kirschneck C, Proff P, Maurer M, Reicheneder C, Römer P. Orthodontic forces add to nicotine-induced loss of periodontal bone: an in vivo and in vitro study. J Orofac Orthop. 2015;76:195–212.CrossRefGoogle Scholar
  8. 8.
    Kirschneck C, Batschkus S, Proff P, Köstler J, Spanier G, Schröder A. Valid gene expression normalization by RT-qPCR in studies on hPDL fibroblasts with focus on orthodontic tooth movement and periodontitis. Sci Rep. 2017;7:14751.CrossRefGoogle Scholar
  9. 9.
    Krishnan V, Nair VS, Ranjith A, Davidovitch Z. Research in tooth movement biology: the current status. Sem Orthod. 2012;18:308–16.CrossRefGoogle Scholar
  10. 10.
    Pilon JJ, Kuijpers-Jagtman AM, Maltha JC. Magnitude of orthodontic forces and rate of bodily tooth movement. An experimental study. Am J Orthod Dentofac Orthop. 1996;110:16–23.CrossRefGoogle Scholar
  11. 11.
    van Leeuwen EJ, Maltha JC, Kuijpers-Jagtman AM. Tooth movement with light continuous and discontinuous forces in beagle dogs. Eur J Oral Sci. 1999;107:468–74.CrossRefGoogle Scholar
  12. 12.
    Giannopoulou C, Dudic A, Pandis N, Kiliaridis S. Slow and fast orthodontic tooth movement: an experimental study on humans. Eur J Orthod. 2016;38:404–8.CrossRefGoogle Scholar
  13. 13.
    Li B, Zhang YH, Wang LX, Li X, Zhang XD. Expression of OPG, RANKL, and RUNX2 in rabbit periodontium under orthodontic force. Genet Mol Res. 2015;14:19382–8.CrossRefGoogle Scholar
  14. 14.
    Shoji-Matsunaga A, Ono T, Hayashi M, Takayanagi H, Moriyama K, Nakashima T. Osteocyte regulation of orthodontic force-mediated tooth movement via RANKL expression. Sci Rep. 2017;18(7):8753.CrossRefGoogle Scholar
  15. 15.
    Fleissig O, Reichenberg E, Tal M, Redlich M, Barkana I, Palmon A. Morphologic and gene expression analysis of periodontal ligament fibroblasts subjected to pressure. Am J Orthod Dentofac Orthop. 2018;154:664–76.CrossRefGoogle Scholar
  16. 16.
    Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, Tomoyasu A, Yano K, Goto M, Murakami A, Tsuda E, Morinaga T, Higashio K, Udagawa N, Takahashi N, Suda T. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci USA. 1998;95:3597–602.CrossRefGoogle Scholar
  17. 17.
    Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003;423:337–42.CrossRefGoogle Scholar
  18. 18.
    Yang CY, Jeon HH, Alshabab A, Lee YJ, Chung CH, Graves DT. RANKL deletion in periodontal ligament and bone lining cells blocks orthodontic tooth movement. Int J Oral Sci. 2018;10:3.CrossRefGoogle Scholar
  19. 19.
    Ishimi Y, Miyaura C, Jin CH, Akatsu T, Abe E, Nakamura Y, Yamaguchi A, Yoshiki S, Matsuda T, Hirano T. IL6 is produced by osteoblasts and induces bone resorption. J Immunol. 1990;145:3297–303.PubMedGoogle Scholar
  20. 20.
    Madureira DF, Taddei Sde A, Abreu MH, Pretti H, Lages EM, da Silva TA. Kinetics of interleukin-6 and chemokine ligands 2 and 3 expression of periodontal tissues during orthodontic tooth movement. Am J Orthod Dentofac Orthop. 2012;142:494–500.CrossRefGoogle Scholar
  21. 21.
    Ranade K, Chang MS, Ting CT, Pei D, Hsiao CF, Olivier M, Pesich R, Hebert J, Chen YD, Dzau VJ, Curb D, Olshen R, Risch N, Cox DR, Botstein D. High-throughput genotyping with single nucleotide polymorphisms. Genome Res. 2001;11:1262–8.PubMedPubMedCentralGoogle Scholar
  22. 22.
    Cunha A, Nelson-Filho P, Marañón-Vásquez GA, Ramos AGC, Dantas B, Sebastiani AM, Silvério F, Omori MA, Rodrigues AS, Teixeira EC, Levy SC, Araújo MC, Matsumoto MAN, Romano FL, Antunes LAA, Costa DJD, Scariot R, Antunes LS, Vieira AR, Küchler EC. Genetic variants in ACTN3 and MYO1H are associated with sagittal and vertical craniofacial skeletal patterns. Arch Oral Biol. 2019;97:85–90.CrossRefGoogle Scholar
  23. 23.
    Giacomini KM, Brett CM, Altman RB, Benowitz NL, Dolan ME, Flockhart DA, Johnson JA, Hayes DF, Klein T, Krauss RM, Kroetz DL, McLeod HL, Nguyen AT, Ratain MJ, Relling MV, Reus V, Roden DM, Schaefer CA, Shuldiner AR, Skaar T, Tantisira K, Tyndale RF, Wang L, Weinshilboum RM, Weiss ST, Zineh I. Pharmacogenetics research network. The pharmacogenetics research network: from SNP discovery to clinical drug response. Clin Pharmacol Ther. 2007;81:328–45.CrossRefGoogle Scholar
  24. 24.
    Frazer KA, Murray SS, Schork NJ, Topol EJ. Human genetic variation and its contribution to complex traits. Nat Rev Genet. 2009;10:241–51.CrossRefGoogle Scholar
  25. 25.
    Mabuchi R, Matsuzaka K, Shimono M. Cell proliferation and cell death in periodontal ligaments during orthodontic tooth movement. J Periodontal Res. 2002;37:118–24.CrossRefGoogle Scholar
  26. 26.
    Pavlin D, Gluhak-Heinrich J. Effect of mechanical loading on periodontal cells. Crit Rev Oral Biol Med. 2001;12:414–24.CrossRefGoogle Scholar
  27. 27.
    Yucel-Lindberg T, Båge T. Inflammatory mediators in the pathogenesis of periodontitis. Expert Rev Mol Med. 2013;15:e7.CrossRefGoogle Scholar
  28. 28.
    Kudo O, Sabokbar A, Pocock A, Itonaga I, Fujikawa Y, Athanasou NA. Interleukin-6 and interleukin-11 support human osteoclast formation by a RANKL-independent mechanism. Bone. 2003;32:1–7.CrossRefGoogle Scholar
  29. 29.
    Chorley BN, Wang X, Campbell MR, Pittman GS, Noureddine MA, Bell DA. Discovery and verification of functional single nucleotide polymorphisms in regulatory genomic regions: current and developing technologies. Mutat Res. 2008;659:147–57.CrossRefGoogle Scholar
  30. 30.
    Sadee W, Wang D, Papp AC, Pinsonneault JK, Smith RM, Moyer RA, Johnson AD. Pharmacogenomics of the RNA world: structural RNA polymorphisms in drug therapy. Clin Pharmacol Ther. 2011;89:355–65.CrossRefGoogle Scholar
  31. 31.
    Tokuyama N, Tanaka S. Cytokines in bone diseases. Genetic disorders of RANKL-RANK-OPG system. Clin Calcium. 2010;20:1532–8.PubMedGoogle Scholar
  32. 32.
    Thirunavukkarasu K, Halladay DL, Miles RR, Yang X. The osteoblast-specific transcription factor Cbfa1 contributes to the expression of osteoprotegerin, a potent inhibitor of osteoclast differentiation and function. J Biol Chem. 2000;275:25163–72.CrossRefGoogle Scholar
  33. 33.
    Lee WC. Experimental study of the effect of prostaglandin administration on tooth movement—with particular emphasis on the relationship to the method of PGE1 administration. Am J Orthod Dentofac Orthop. 1990;98:231–41.CrossRefGoogle Scholar
  34. 34.
    Albert PR. What is a functional genetic polymorphism? Defining classes of functionality. J Psychiatry Neurosci. 2011;36:363–5.CrossRefGoogle Scholar
  35. 35.
    Nettelhoff L, Grimm S, Jacobs C, Walter C, Pabst AM, Goldschmitt J, Wehrbein H. Influence of mechanical compression on human periodontal ligament fibroblasts and osteoblasts. Clin Oral Investig. 2016;20:621–9.CrossRefGoogle Scholar

Copyright information

© The Society of The Nippon Dental University 2019

Authors and Affiliations

  1. 1.Department of Pediatric Dentistry, School of Dentistry of Ribeirão PretoUniversidade de São PauloRibeirão PretoBrazil
  2. 2.Department of OrthodonticsUniversity of RegensburgRegensburgGermany
  3. 3.Department of Oral and Maxillofacial SurgeryUniversidade PositivoCuritibaBrazil
  4. 4.Department of Cranial and Maxillofacial SurgeryUniversity of RegensburgRegensburgGermany

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